干扰素诱导跨膜蛋白1对结肠癌SW480细胞株侵袭和转移能力的影响

Influences of the interferon induced transmembrane protein 1 on the invasion and metastasis of colorectal cancer SW480 cell lines

目的研究干扰素诱导跨膜蛋白1(IFITM1)对结肠癌SW480细胞株侵袭和转移的影响。方法构建IF-ITM1/pEGFP-C3重组质粒并且转染大肠癌细胞株SW480,免疫荧光显微镜和激光共聚焦显微镜(LCSM)、RT-PCR和Westernblotting鉴定IFITM1/pEGFP-C3重组质粒。G418筛选稳定过表达IFITM1的SW480细胞株(IFITM1/SW480),细胞侵袭性实验、明胶酶法测定MMP-2和MMP-9活性,Western blotting检测MMP-9蛋白表达及IFITM1/SW480侵袭和转移能力。结果成功构建IFITM1/pEGFP-C3重组表达质粒,获得IFITM1/SW480细胞株;细胞侵袭实验显示SW480细胞、IFITM1/SW480细胞的细胞穿透数分别为(448.64±38.09)个和(540.45±44.61)个,差异有统计学意义(P<0.01);明胶酶法测定显示IFITM1/SW480细胞MMP-2、MMP-9活性较SW480和pEGPF/SW480细胞明显增强。Western blottin显示IFITM1/SW480细胞MMP-9表达水平明显高于SW480和pEGFP/SW480细胞株。结论 IFITM1可增加大肠癌SW480细胞株侵袭和转移的能力。

Objective To study the influences of the interferon-induced transmembrane protein 1（IFITM1） on the invasion and metastasis of colorectal cancer SW480 cell lines. Methods We constructed IFITM1/pEGFP-C3 recombinant plasmids and transfected them into the colorectal cancer SW480 cell lines.IFITM1/pEGFP-C3 recombinant plasmids were identified by means of immunofluorescence,laser confocal scanning microscopy,reverse transcription polymerase chain reaction（RT-PCR）and Western blotting.IFITM1/SW480 cells with stable over-expression of IFITM1 were confirmed by G418 screening.Cell invasion experiments were performed to determine the invasion capacity of the IFITM1/SW480 cells.MMP-2 and MMP-9 activities were detected by the gelatin zymographic analysis,and MMP-9 expression was detected by the Western blotting analysis. Results IFITM1/pEGFP-C3 recombinant plasmids were successfully constructed in this study,and the IFITM1/SW480 cells with stable IFITM1 gene over-expression was confirmed by G418 screening.The numbers of the SW480 cells and IFITM1/SW480 cells migrated through matrigel were 448.64±38.09 and 540.45±44.61 with statistic significance（P0.01）.Gelatin zymographic analysis showed that MMP-9 and MMP-2 activities in the IFITM1/SW480 cells were significantly enhanced,and Western blotting analysis showed that MMP-9 expression in the IFITM1/SW480 cells was also increased compared with SW480 and pEGFP/SW480 cell lines. Conclusion IFITM1 can enhance the invasion and metastasis of the colorectal cancer SW480 cell lines.